Method for measuring chemiluminescence

• US 9,869,672 B2
• Filed: 03/19/2013
• Issued: 01/16/2018
• Est. Priority Date: 03/19/2012
• Status: Active Grant

• Current Assignee: Jyvaskylan Yliopisto
• Original Assignee: Jyvaskylan Yliopisto
• Inventors: Tiirola, Marja
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US14/382,512
• Publication Number:
  US 20150065370A1
• Time in Patent Office: 1,764 Days
• Field of Search: None
• CPC Class Codes: G01N 27/4145 specially adapted for biomolecules, e.g. gate electrode with immobilised receptors
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/60 involving radioactive labelled substances
• CPC Class Codes: G03C 5/26 Processes using silver-salt-containing photosensitive materials or agents therefor physical development G03C5/58

Abstract

The present invention provides a method for determining radioactivity in an ion-sensitive field effect transistor array, the method comprising the steps of incubating the array with electron-sensitive silver-halide material and then incubating the array with a developer solution reducing the silver halides that have been exposed to radioactivity to elemental silver and simultaneously measuring pH at each separate reaction chambers, wherein decrease of the pH indicates the presence of radioactivity in a reaction chamber.

Claims

1. Method for determining chemiluminescence in an ion-sensitive field effect transistor array comprising the steps of:

a) attaching molecules of a sample to reaction chambers in said array;

b) contacting said array with a chemiluminescence causing enzymatic molecule conjugated to a molecule with specific binding properties to said molecules;

c) incubating the array with light-sensitive silver-halide material and a chemiluminescence substrate to produce chemiluminescence light thereby exposing the light-sensitive silver-halide material to chemiluminescent light;

d) incubating the array with a developer solution reducing the silver halides that have been exposed to the chemiluminescence light to elemental silver thereby forming or enhancing a latent image; and

e) determining concentration of the chemiluminescence causing enzymatic molecule by measuring a decrease in pH at each separate reaction chamber, said decrease in pH resulting from latent image formation or latent image enhancement.

2. The method according to claim 1, wherein the chemiluminescence causing enzymatic molecule is selected from the group consisting of horseradish peroxidase, alkaline phosphatase and luciferase.

3. The method according to claim 1, wherein the chemiluminescence substrate is selected from the group consisting of luminol, dioxetane, acridinium ester, hydrazide and luciferin.

4. The method according to claim 1, wherein the developer solution is an unbuffered solution or a solution with a low buffer capacity that has a pH 10-11.

5. The method according to claim 1, wherein the developer solution is an ascorbic acid based unbuffered developer solution.

6. The method according to claim 5, wherein the ascorbic acid based unbuffered developer solution comprises 1-phenyl-3-pyrazolidon.